Research internship on biotechnology-assisted varietal development in pineapple

Date

12-2014

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Hayde F. Galvez

Co-adviser

Lolita D. Valencia

Abstract

The research internship was conducted at the Crop Science Cluster-Institute of Plant Breeding, College of Agriculture, University of the Philippines Los Baños from April 2014 to December 2014. The wet laboratory and field exposure were conducted at the Fruit and Ornamental Crops Breeding Section. While the second part of the research internship , which involved basic molecular biocomputing analysis, was conducted at the Genetics Laboratory. Queen and Smooth Cayenne pineapple cultivars were induced to flowering using Ethrel(ᴿ) SL 480 solution supplemented with urea. To be trained on pollen viability and germinability assays, 75% and 100% (fully) opened florets were used. The aceto-carmine (2%) assay was used and pollen examination was done using the Reichert-Jung binocular light microscope at 400x magnification. For further analysis, pollen grains were also collected and stored in the laboratory. To generate seeds and be skilled in pineapple hybridization, varietal crosses were performed among seven(7) varieties between Smooth Cayenne and Queen pineapple cultivars. With another set of fruiting pineapple plants, pineapple fruits were harvested and evaluated for fruit characteristics and morphology. Seeds from hybrid fruits were extracted and properly stored. The percentage (%) of seed germination was also evaluated using the fine-sand paper scarification method vs. without scarification. Using appropriate bioinformatics tools, PCR primers were designed to target and amplify specific genome regions of some genes involved in fruit ripening and color development. The genes characterized were phytoene desaturase, phythoene synthase and carotene hydroxylase. Web-based bioinformatics tools used included access to the Sol Genomics Network (SGN), specifically under Tomato database, and NCBI databases, and the GBrowse, Primer3 and Primer-BLAST programs. The specificity and comparative genomics application of the PCR primer designed was validated by Primer-BLAST search in the NCBI database. After further manual validation of the stringency of the nucleotide sequence parameters, the PCR primers can be synthesized and optimized to amplify the homolog genes in pineapple and other fruit crops.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993 2014 A127 /I23

Notes

major: Crop Biotechnology

Document Type

Thesis

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